The overall objective of this project is to study the dietary and hormonal regulation of fatty acid metabolism. Diabetes mellitus decreases fatty acid synthesis by decreasing the synthesis of fatty acid synthetase and acetyl-CoA carboxylase. Insulin injection to diabetic animals or 17 day chick embryos induces fatty acid synthetase. We plan to study the role of insulin and related hormones on the synthesis of fatty acid synthetase and acetyl-CoA carboxylase in intact animals and in cultured cells. We have recently isolated the fatty acid synthetase mRNA from the uropygial gland of goose lactating rat mammary gland and insulin induced chick embryo livers. Translation of the isolated mRNAs in a cell free system yield proteins which have the same as the native synthetase subunits and are immunologically identical to their respective native synthetases. Attempts are underway to prepare the cDNAs and insert them into appropriate host cells. Clones carrying the cDNA plasmids will be identified and utilized in the preparation of large scale cDNA. The latter will be used in studying the effect of insulin on the formation and stability of the mRNA and its translation by the ribosomes will be studied. The effect of insulin, cortisol and prolactin on the induction of fatty acid synthetase and the lipogenic enzymes in mammary gland explants will be investigated. Methodology to be employed in the outlined studies include enzymology, protein chemistry, lipid chemistry and cell biology.